Background

NIH Common Fund Extracellular RNA Communication program (https://commonfund.nih.gov/exrna) was established in 2012 to uncover fundamental principles of exRNA generation, secretion, and transport; identify and develop a catalog of exRNA found in normal human body fluids; and investigate the potential for using exRNAs in the clinic as therapeutic molecules and/or biomarkers of disease. The Common Fund supports cross-cutting programs that are expected to have exceptionally high impact. All Common Fund initiatives invite investigators to develop bold, innovative, and often risky approaches to address problems that may seem intractable or to seize new opportunities that offer the potential for transformation of research processes.

ExRNAs can act as endocrine signaling molecules, both locally and systemically, representing a novel paradigm in intercellular communication. ExRNAs are transported in body fluids in association with an array of carrier vehicles of varying complexity including extracellular vesicles (EVs), ribonucleoproteins (RNPs), and lipoproteins (LPPs). While these carriers (EVs, RBPs, and LPPs) share the functional attribute of protecting exRNAs from degradation by ubiquitous extracellular RNAses, their respective contributions to biodistribution, uptake, and/or function of their respective exRNA cargo in target cells is less clear. In humans, exRNAs have been found in virtually all body fluids examined, including blood, saliva, urine, breast milk, cerebral spinal fluid (CSF), amniotic fluid, semen, ascites, and pleural effusions. Reports in the literature suggest that exRNAs can have both protective and pathogenic roles in a variety of human diseases, and concomitantly serve as biomarkers for disease states. Taken together, the above findings highlight the important role secreted exRNAs have in regulating human health and disease.

While significant strides have been made in the field of exRNA biology, specific technological challenges remain. For example, RNA concentrations in biofluids are significantly lower than in tissues, necessitating the development of improved methods for their isolation and analysis. Currently available separation approaches do not adequately discriminate exRNAs encapsulated in EVs from exRNAs associated with non-vesicular RNPs or LPPs, and the carriers that transport exRNAs can affect exRNA recovery when different isolation methods are used. Ultimately, this can lead to unintended biases in exRNA population profiling. Little is known about the normal physiology of exRNAs, the functional consequences of exRNA uptake, or deficiency of uptake, by recipient cells, and the level of variation existing at the subpopulation level in both normal and disease states. These technological impediments limit the utility and impact of current efforts to catalog exRNAs and assess their contribution(s) to human health and disease.

These technology challenges will be addressed in the second stage of the NIH Common Fund exRNA Communication Program Advancing Extracellular RNA (exRNA) Communication Research. Three companion FOAs will focus on the development of tools, technologies, and resources to catalyze the field of exRNA communication research.

• RFA-RM-18-026: Limited Competition: Data Management and Resource Repository (DMRR) on Extracellular RNA (U54 Clinical Trial Not Allowed). This companion FOA will support the DMRR to curate and disseminate information regarding critical reagents and resources, as well as promote and facilitate cross-project collaborations
• RFA-RM-18-027: Advancing Extracellular RNA (exRNA) Communication Research: Improved Isolation and Analysis of exRNA-Carrier Subclasses (UG3/UH3 Clinical Trial Not Allowed). This companion FOA will specifically focus on improved separation technologies to rapidly and reproducibly sort exRNA carrier vehicles (EVs, RBPs, and LPPs) and identify their respective exRNAomes and associated molecular cargo.
• RFA-RM-18-028: Advancing Extracellular RNA (exRNA) Communication Research: Towards Single Extracellular Vesicle (EV) Sorting, Isolation, and Analysis of Cargo (UG3/UH3 Clinical Trial Not Allowed). This FOA will specifically focus on single EV isolation and analysis tools to assess exRNA heterogeneity and enable mapping exRNAs to their cell or tissue of origin. A full description of this goals of this FOA is below in Research Objectives.

Development of these tools, technologies, and resources will complement Stage 1 products and fill remaining knowledge and technology gaps. A consortium-structured grant mechanism is being used to support this research initiative to facilitate the generation of community accepted standards. Meeting the goals of the Stage 2 Initiatives will have a transformative effect on the field of exRNA biology as this will enable a greater understanding of the fundamental role of exRNAs in intercellular signaling and the translational potential to diagnose and treat diseases.

Research Objectives

Extracellular vesicles (EVs) are membrane-enclosed particles that are secreted from various cell types into the extracellular space. EVs are heterogeneous in size, origin, and molecular cargo (including proteins, RNA, DNAs, and lipids). Additional complexity exists due to the heterogeneous exRNAs (mRNA, miRNA, lncRNA, and other RNA species) within EVs. These exRNAs can be transported to distal sites and may play an important role in intercellular communication by regulating physiological and pathological processes. Current methods for isolating EVs from complex biofluids are limited by the inability to sort EVs by cell or tissue of origin, creating significant technological barriers to analytical comparison of datasets generated across studies. Currently, it is not clear which RNA species (if any) are packaged and transported by different EV sub-populations, if the exRNA carrying capacity of different EV populations differs, or whether the exRNA content of EVs differs under homeostatic or pathologic conditions. Further, current technologies do not permit single EV isolation and analysis thereby limiting our understanding of the normal variation (heterogeneity) of EVs produced even using reductionist (i.e., cell culture) models. The diagnostic or therapeutic functionality of exRNAs can only be truly realized once the range of EV subpopulations from a given cell source are fully described and characterized for complete analysis of molecular cargo. Determining the origin of EVs from various tissues or cell types is essential for understanding where their cargo exRNAs originate and their effect on their target cells. Furthermore, being able to isolate and analyze single EVs from human biofluids would provide a unique opportunity to understand the cell or tissue from which their respective exRNAs originate and, importantly, would add significant depth to our understanding of exRNA communication. Ideally, a protocol(s) for isolating single vesicles should be (1) rapid and reproducible, (2) yield highly enriched EV subpopulations from a single source cell or tissue with no cross-contamination, and (3) have the capacity to analyze and catalog exRNAs present in the single EVs from various cell and tissue types. None of the separation technologies currently employed for EVs (i.e., differential ultracentrifugation, density gradients, polymer-based precipitation, microfiltration and size-exclusion-based methods) meet all these performance standards. Further, the diagnostic or therapeutic functionality of EVs can only be truly realized once the range of EV subpopulations from a given cell source are fully described and isolated for complete analysis of their molecular cargo. A number of approaches have been developed to characterize EVs in the clinical setting, including sandwich ELISA-based microarray chip, immunomagnetic exosome RNA (iMER) analysis, miniaturized micro nuclear magnetic resonance ( NMR) microfluidic chip system, Exochip, and label-free high-throughput nano-plasmonic exosome assay (nPLEX) using surface plasmon resonance (SPR). However, none of these current methodologies can address EV heterogeneity or clearly define cell of origin for exRNA cargo.

Thus, there is a critical need to develop and validate novel tools, technologies, and approaches to characterize and sort EV subpopulations based on cell of origin from complex biofluids for downstream analysis and cataloging of their respective exRNAs and associated molecular cargo, with the capacity to do so at scale.

This FOA uses the UG3/UH3 cooperative agreement award mechanism. The UG3 phase will focus on technology development and feasibility using specific human cultured cells or tissue types (primary derived biopsies), animal models, or human biofluids. Support will be provided for up to two years for the initial proof of concept studies. Following NIH administrative review, the UH3 Phase will support activities to optimize, refine, and scale up validated EV isolation technologies, and using human biofluids. Successful technologies, information on PD/PI generated reagents, and associated protocols will be shared with the broader community via the exRNA Portal (exRNA.org). This will be the primary deliverable of this collaborative Common Fund-sponsored program.

In order to be responsive to this FOA, applicants are expected to:

• Propose strategy/strategies for isolating and tracking EV subpopulations, cataloging EV-associated exRNAs based on cell or tissue of origin and intended target cell. Applications aiming to characterize exRNAs associated with other carrier vehicles (RBPs or LPPs) should apply to RFA-RM18-027.
• Outline strategy/strategies that allow for high-throughput separation and analysis of EV subpopulation-specific molecular cargo including genomics, proteomics, and lipidomics.
• Justify how the proposed EV isolation strategy/strategies will improve the state of the art either by enabling purification of single EVs in a way that has previously not been possible, or will dramatically improve (by at least 5-fold) our ability to purify EV subpopulations with respect to increased purity, increased speed, or yield.
• Demonstrate isolation effectiveness using a variety of human biofluids in the UH3 Phase.
• Incorporate approaches to share the strategies/tools developed with the broader scientific community.

UG3/UH3 transitions and milestones:

All projects will be milestone-driven with clear go/no-go quantifiable criteria. Prior to funding an application, the Program Official will contact the applicant to discuss the proposed UG3 and UH3 milestones and any changes suggested by NIH staff or the NIH review panel. The Program Official and the applicant will negotiate and agree on a final set of approved UG3 milestones which will be specified in the Notice of Award. These milestones will be the basis for judging the successful completion of the work proposed in the UG3 stage and progress towards interim milestones in the UH3 stage. Only UG3 projects that meet their milestones will have an opportunity to move to the UH3 phase. UH3 milestones will be the basis for judging progress towards and completion of interim milestones in the UH3 stage.

Applicants are expected to carefully read Section IV 2 for detailed application instructions.

Program Formation and Governance

The awards funded under this FOA will be cooperative agreements (see Section VI. 2. Cooperative Agreement Terms and Conditions of Award). Close interactions among the awardees and NIH will be required to maintain this complex program. The exRNA Communication Consortium (ERCC) governance will rest with the ERCC Program Steering Committee in collaboration with NIH Program Officials, with advice from a group of External Program Consultants (EPC) providing critical scientific and managerial insights, and subject to oversight by the NIH ERCC Working Group. The NIH ERCC Working Group consists of NIH programmatic staff from multiple Institutes and Centers of the NIH and the NIH Office of the Director. This group will be primarily responsible for the stewardship of the ERCC program. The NIH ERCC Working Group is chaired by the Director of the National Center for Advancing Translational Science (NCATS) and Director for the Division of Cancer Biology, National Cancer Institute (NCI). It reports to the Directors of the Office of Strategic Coordination/Common Fund and the Division of Program Coordination, Planning, and Strategic Initiatives for final funding decisions.

By participating in the exRNA Communication Consortium, the awardees are expected to agree to:

Communication Plan

• Participate in regular (monthly) conference calls with all consortium members
• Coordinate efforts with other awardees, in particular with the DMRR
• Participate and present findings at meetings convened by the NIH through the DMRR twice a year in the DC area
• Jointly publish findings in a timely manner

Performance Requirements

• Meet yearly milestones as defined by investigators and NIH Program Official at the time of award
• Work with, cooperatively interact with, and actively seek input from NIH

Sharing Plan

• Broad sharing of data and biological specimens with academic, industry, and government researchers is a critical feature of this program that is consistent with applicable laws, regulations, and policies. All subjects must be properly consented to allow appropriate sample and data distribution to researchers in academia and industry.

The DMRR and designated NIH databases will be utilized for the banking and distribution of biological and clinical data. Project PD(s)/PI(s) will be expected to harmonize all data and resource generation with the Data Management and Resource Repository (DMRR). The DMRR is expected to curate and disseminate information regarding critical reagents, and resources and promote and facilitate cross-project collaborations (see RFA-RM-18-026).

Technical Assistance Webinar

All applicants are strongly encouraged to contact NIH staff to discuss the alignment of their proposed work with the goals of this FOA and the ERCC program. A technical assistance webinar will be held for potential applicants. NIH staff will be available to answer questions related to this and companion FOAs. Participation in the webinar is optional. No pre-registration is required, but participants are asked to submit questions ahead of time to [email protected]. The slides as well as some frequently asked questions will be available at https://commonfund.nih.gov/exrna following the webinar.

Webinar Information:

The webinar will be held on August 24, 2018 from 2-3:30 PM, EST.

Join us via WebEx using this link:
https://nih.webex.com/nih/onstage/g.php?MTID=ef29317fb2d940f2f6f0c3c410e5df5ff

Call-in toll number (US/Canada): 1-650-479-3208

Event number: 629 822 206

Event password: exRNA

Higher Education Institutions

• Public/State Controlled Institutions of Higher Education
• Private Institutions of Higher Education

The following types of Higher Education Institutions are always encouraged to apply for NIH support as Public or Private Institutions of Higher Education:

• Hispanic-serving Institutions
• Historically Black Colleges and Universities (HBCUs)
• Tribally Controlled Colleges and Universities (TCCUs)
• Alaska Native and Native Hawaiian Serving Institutions
• Asian American Native American Pacific Islander Serving Institutions (AANAPISIs)

Nonprofits Other Than Institutions of Higher Education

• Nonprofits with 501(c)(3) IRS Status (Other than Institutions of Higher Education)
• Nonprofits without 501(c)(3) IRS Status (Other than Institutions of Higher Education)

For-Profit Organizations

• Small Businesses
• For-Profit Organizations (Other than Small Businesses)

Governments

• State Governments
• County Governments
• City or Township Governments
• Special District Governments
• Indian/Native American Tribal Governments (Federally Recognized)
• Indian/Native American Tribal Governments (Other than Federally Recognized)
• Eligible Agencies of the Federal Government, including NIH Intramural program
• U.S. Territory or Possession

Other

• Independent School Districts
• Public Housing Authorities/Indian Housing Authorities
• Native American Tribal Organizations (other than Federally recognized tribal governments)
• Faith-based or Community-based Organizations
• Regional Organizations
• Non-domestic (non-U.S.) Entities (Foreign Institutions)

Applicant organizations

Applicant organizations must complete and maintain the following registrations as described in the SF 424 (R&R) Application Guide to be eligible to apply for or receive an award. All registrations must be completed prior to the application being submitted. Registration can take 6 weeks or more, so applicants should begin the registration process as soon as possible. The NIH Policy on Late Submission of Grant Applications states that failure to complete registrations in advance of a due date is not a valid reason for a late submission.

• Dun and Bradstreet Universal Numbering System (DUNS) - All registrations require that applicants be issued a DUNS number. After obtaining a DUNS number, applicants can begin both SAM and eRA Commons registrations. The same DUNS number must be used for all registrations, as well as on the grant application.
• System for Award Management (SAM) (formerly CCR) Applicants must complete and maintain an active registration, which requires renewal at least annually. The renewal process may require as much time as the initial registration. SAM registration includes the assignment of a Commercial and Government Entity (CAGE) Code for domestic organizations which have not already been assigned a CAGE Code.
  • NATO Commercial and Government Entity (NCAGE) Code Foreign organizations must obtain an NCAGE code (in lieu of a CAGE code) in order to register in SAM.
• eRA Commons - Applicants must have an active DUNS number and SAM registration in order to complete the eRA Commons registration. Organizations can register with the eRA Commons as they are working through their SAM or Grants.gov registration. eRA Commons requires organizations to identify at least one Signing Official (SO) and at least one Program Director/Principal Investigator (PD/PI) account in order to submit an application.
• Grants.gov Applicants must have an active DUNS number and SAM registration in order to complete the Grants.gov registration.

Program Directors/Principal Investigators (PD(s)/PI(s))

All PD(s)/PI(s) must have an eRA Commons account. PD(s)/PI(s) should work with their organizational officials to either create a new account or to affiliate their existing account with the applicant organization in eRA Commons. If the PD/PI is also the organizational Signing Official, they must have two distinct eRA Commons accounts, one for each role. Obtaining an eRA Commons account can take up to 2 weeks.

Any individual(s) with the skills, knowledge, and resources necessary to carry out the proposed research as the Program Director(s)/Principal Investigator(s) (PD(s)/PI(s)) is invited to work with his/her organization to develop an application for support. Individuals from underrepresented racial and ethnic groups as well as individuals with disabilities are always encouraged to apply for NIH support.

For institutions/organizations proposing multiple PDs/PIs, visit the Multiple Program Director/Principal Investigator Policy and submission details in the Senior/Key Person Profile (Expanded) Component of the SF424 (R&R) Application Guide.

Applicant organizations may submit more than one application, provided that each application is scientifically distinct.

The NIH will not accept duplicate or highly overlapping applications under review at the same time. This means that the NIH will not accept:

• A new (A0) application that is submitted before issuance of the summary statement from the review of an overlapping new (A0) or resubmission (A1) application.
• A resubmission (A1) application that is submitted before issuance of the summary statement from the review of the previous new (A0) application.
• An application that has substantial overlap with another application pending appeal of initial peer review (see NOT-OD-11-101)

All page limitations described in the SF424 Application Guide and the Table of Page Limits must be followed. The following section supplements the instructions found in the SF424 (R&R) Application Guide and should be used for preparing an application to this FOA. All instructions in the SF424 (R&R) Application Guide must be followed. All instructions in the SF424 (R&R) Application Guide must be followed. All instructions in the SF424 (R&R) Application Guide must be followed. All instructions in the SF424 (R&R) Application Guide must be followed. All instructions in the SF424 (R&R) Application Guide must be followed. All instructions in the SF424 (R&R) Application Guide must be followed. All instructions in the SF424 (R&R) Application Guide must be followed, with the following additional instructions:

Specific Aims:In the single Specific Aims attachment, provide the overall goals or hypotheses for the entire project period and identify separate Specific Aims to be accomplished in the UG3 phase and in the UH3 phase.

Research Strategy:

The UG3/UH3 phases must be submitted as a single application but the approach should be clearly organized into two stages: UG3 Development (phase 1) and UH3 Optimization (phase 2). All projects should be milestone-driven with clear go/no-go criteria that are quantifiable. Milestones and the timeline for each stage must be provided in a separate heading at the end of the Approach section for each UG3 and UH3 component and include the following:

• Provide detailed quantitative criteria by which milestone achievement will be assessed.
• Provide a detailed timeline (Gantt chart) for the anticipated attainment of each milestone and the overall goal.
• Yearly quantitative milestones are required to provide clear indicators of a project’s continued success or emergent difficulties.
• Identify any impediments that could require an addendum to the research plan, milestones, or timeline with a discussion of alternative approaches.
• Include a clearly identified Go/No-Go transition milestone for completion of the UG3 phase at the end of Year 2 and transition to the UH3 phase for 2 years of additional funding.

Appropriate benchmarks for assessing milestones and project transitions will be negotiated between the NIH and applicant prior to funding decisions and will be specified in the Notice of Award.

The UG3 Development Phase:

UG3 projects will support feasibility and proof of principle studies for novel isolation technologies for EV subpopulations using physiologically relevant human 3D cell models, tissue types derived from primary biopsies, animal models, or human biofluids. Studies will focus on identifying and cataloging exRNA cargos in identified EV subpopulations in order to characterize cell of origin and intended target cell. Molecular tools as well as genetic or barcoding-based systems that allow assessment of separation performance (i.e., recovery, purity, functional output) of specific EV subpopulations are also encouraged. The emphasis of the UG3 phase is to develop separation technologies capable of rapidly separating EV subpopulations into relatively homogenous populations (based on biophysical characteristics) from a single sample. UG3 investigators will need to be flexible in the design and execution of their specific applications through collaboration with other UG3 groups in identifying the optimum protocol(s) for isolating EV subpopulations from various biofluids or tissue types, and to develop common best practices. Applicants should:

• Provide a description of the hypothesis to be tested in the UG3 phase of the study
• Provide a description of a protocol(s) for isolating single vesicles that is (1) rapid and reproducible, (2) yields highly enriched EV subpopulations from a single source cell or tissue with no cross-contamination, (3) has the capacity to analyze and catalog exRNAs present in the single EVs from various cell and tissue types (4) can track EV subpopulations, and (5) catalogs EV-associated exRNAs based on cell or tissue of origin and intended target cell.
• Include an approach that allows for high-throughput separation and analysis of EV subpopulation-specific molecular cargo including genomics, proteomics, and lipidomics.
• Justify how the proposed EV isolation strategy/strategies will improve the state of the art, either by enabling purification of single EVs in a way that has previously not been possible, or will dramatically improve (by at least 5-fold) our ability to purify EV subpopulations with respect to increased purity, increased speed, or yield.
• Incorporate approaches to share the strategies/tools developed with the broader scientific community via the exRNA Portal (exRNA.org).

Transition to the UH3 Optimization Phase:

UG3 projects that have met the scientific milestones and other requirements (listed below) will be eligible for transition to the second UH3 Optimization Phase after NIH administrative review.

• Successful achievement of the defined milestones (including go/no-go conditions) for the UG3 Development Phase of the project.
• Potential for meeting the goal of moving towards single-EV characterization.
• Evidence of having collaborated with other awardees within and outside a Consortium arrangement using the set-aside collaborative budget to meet the goals of the program; interacting with the DMRR; work with the outside community and disseminate resources or tools generated in an expedient manner.
• Availability of funds.
• Program priorities.
• Consistent with recent NIH guidance on rigor and reproducibility in research (https://grants.nih.gov/reproducibility/index.htm), the most important milestone for the transition will be independent validation of any separation tools/technologies/approaches. To achieve this milestone, each applicant must set-aside approximately 10% (Direct Costs) of year 2-4 funds to establish a collaboration(s) with a group(s) who will independently validate and provide an assessment of the potential utility of the tools/technologies/approaches.
• These awards will be assessed and renewed on an annual basis, with renewal contingent on successful attainment of goals and milestones.

Go/No-Go Transition Milestone for transition from the UG3 Phase to the UH3 Phase:

Applicants must be supported by clearly identified Go/No-Go transition milestones for completion of the UG3 phase at the end of Year 2 and transition to the UH3 phase and timelines for assessing progress.

UH3 Optimization Phase:

The UH3 Optimization Phase should focus on optimizing, refinement, and scale up validated EV isolation technologies, towards the end goal of the analysis of single EVs isolated from complex human biofluids. Project in the UH3 phase must include the use of human biofluids. Resulting data should be compared to previously deposited exRNA Atlas datasets (http://exrna-atlas.org/) with the goal of determining the exRNA cell of origin and intended target cell. Activities supported in this phase include, but are not limited to:

• Optimizing tools/technologies/approaches developed during the UG3 Phase using multiple human biofluids.
• Establishing a catalog of exRNAs present in complex human biofluids, their respective cell of origin, and intended target cell.
• Identifying biophysical attributes capable of improving separation of single EVs.
• Creating scale-up of process development, formulation and stability studies, using cGMP-compatible methodologies.

Demonstrating isolation effectiveness using a variety of human biofluids in the UH3 Phase.

• All applications, regardless of the amount of direct costs requested for any one year, should address a Data Sharing Plan.
• Applicants should indicate their willingness to abide by all data deposition, quality control metrics, standardization, metadata requirements, data and software release, and public copyright license policies developed by the exRNA Program Steering Committee and approved by NIH staff. A primary goal of the exRNA program is to facilitate discoveries by the broad scientific community. Restrictive licensing terms and sharing practices for exRNA-generated data, tools, and resources could substantially diminish their value and public benefit. Accordingly, awardees should manage data, resources, protocols, tools, and software in a way that achieves this goal. Sharing practices that would hinder, prevent or block access to or use of exRNA program data, tools, and resources for research purposes will be considered to be hindering the goals of the exRNA program. The development of policies, methods, and standards for such sharing is critically important. The NIH expects that the awardees, through the exRNA Program Steering Committee, will develop such policies, methods, and standards in concert with the NIH. These policies, methods, and standards will remain consistent with NIH-wide policies on data and resource sharing.
• Specific Plan for Data Sharing: Consistent with achieving the goals of this program, the NIH expects that information such as collected data (e.g. metabolomic, lipidomic, proteomic, etc.), technical protocols, and any other data or metadata collected under this FOA is to be deposited as appropriate into existing, publicly available data repositories, in particular the DMRR, that are easily accessible, and in machine readable format. Applicants must adhere to FAIR data principles (Findable, Accessible, Interoperable, and Reusable), data standards and processes to be determined by the NIH Data Commons, and evolving NIH data policies. Where appropriate, applicants should identify such repositories and plans for deposition. For datatypes that lack suitable public repositories, applicants should indicate their willingness to identify an appropriate alternative solution that is consistent with achieving the goals of the program. Data should also be made available as appropriate via the exRNA Portal that will serve as the central access point for information regarding data, critical tools, protocols and reagents being developed by the exRNA program.
• Genomic Data Sharing Plan: If applicants propose to generate transcriptomic or other types of genomic data, they must indicate their willingness to abide by the NIH Genomic Data Sharing Policy (https://gds.nih.gov/) and should indicate their agreement to it in the data sharing plan.
• Specific Plan for Protocol, Tool, and Reagent Sharing: As one of the primary goals of this program is to advance research through development, establishment, broad dissemination and use of community resources across the research community, NIH intends that protocols, tools, and reagents generated by the exRNA program be broadly available and distributed at minimal cost, and without undue intellectual property constraints, so that they can be as widely used as possible for research purposes by the larger scientific community. For all applications and where otherwise applicable, the applicant should discuss plans for sharing and distribution of non-data resources that will be generated by the proposed project, including protocols, instrument design specifications, animal strains, plasmids, viruses, antibodies, chemical compounds, biomaterials, and other reagents.
• The exRNA DMRR will work with all exRNA program investigators to collect, curate, and disseminate information regarding tools and reagents being developed by the program and to disseminate this information through the exRNA Portal and other sources as appropriate and consistent with achieving the goals of the program.
• Specific Plan for Sharing Software: A software dissemination plan, with appropriate timelines, is expected in applications that are developing software. There is no prescribed single license for software produced in this project; however, reviewers will be asked to evaluate the software sharing and dissemination plan based on its likely impact. A dissemination plan guided by the following principles is thought to promote the largest impact:

• The software should be freely available to biomedical researchers and educators in the non-profit sector, such as institutions of education, research institutions, and government laboratories.
• The terms should also permit the dissemination and commercialization of enhanced or customized versions of the software, or incorporation of the software or pieces of it into other software packages.
• To preserve utility to the community, the software should be transferable such that another individual or team can continue development in the event that the original investigators are unwilling or unable to do so.
• The terms of software availability should include the ability of researchers outside the project and its collaborating projects to modify the source code and to share modifications with other colleagues. An applicant should take responsibility for creating the original and subsequent "official" versions of a piece of software.
• Applicants are asked to propose a plan to manage and disseminate the improvements or customizations of their tools and resources by others. This proposal may include a plan to incorporate the enhancements into the "official" core software, may involve the creation of an infrastructure for plug-ins, or may describe some other solution.
• Any software dissemination plans represent a commitment by the institution (and its subcontractors as applicable) to support and abide by the plan.

• Applicants should also be familiar with the NIH statements regarding intellectual property of resources developed with Federal funds (NIH Research Tools Policy (http://grants.nih.gov/grants/intell-property_64FR72090.pdf) and other related NIH sharing policies at http://sharing.nih.gov).

Sharing within the ERCC

The ERC program aims to maximize the benefits of sharing, while protecting the intellectual property of the data generators. Prepublication sharing of data within the consortium will allow collaboration across initiatives and further the progress of the ERC Program towards its goals and objectives. Therefore, the ERC program requires that data, metadata, and resources generated from ERC funding be made available to other ERCC members immediately upon being considered shareable . When ERC-generated material under embargo is shared within the ERCC, it will be considered confidential with the understanding that the information will not be used or disclosed by ERCC members unless explicitly agreed upon by the originator of the data under mutually acceptable terms. That is, Consortium members may not publish using material generated by other consortium members without a collaboration or other agreement, or until the material in question has been made available to the public . As such, ERCC investigators accessing embargoed material only available to members of the ERCC must sign a non-disclosure form agreeing to these terms.

Prior to funding, NIH Program Staff may negotiate modifications to the Sharing Plan with the applicant. Only limited Appendix materials are allowed. Follow all instructions for the Appendix as described in the SF424 (R&R) Application Guide. When involving NIH-defined human subjects research, clinical research, and/or clinical trials (and when applicable, clinical trials research experience) follow all instructions for the PHS Human Subjects and Clinical Trials Information form in the SF424 (R&R) Application Guide, with the following additional instructions:

If you answered Yes to the question Are Human Subjects Involved? on the R&R Other Project Information form, you must include at least one human subjects study record using the Study Record: PHS Human Subjects and Clinical Trials Information form or Delayed Onset Study record.

Study Record: PHS Human Subjects and Clinical Trials Information

All instructions in the SF424 (R&R) Application Guide must be followed.

All instructions in the SF424 (R&R) Application Guide must be followed. Applicants are required to follow the instructions for post-submission materials, as described in the policy. Any instructions provided here are in addition to the instructions in the policy. Reviewers will provide an overall impact score to reflect their assessment of the likelihood for the project to exert a sustained, powerful influence on the research field(s) involved, in consideration of the following review criteria and additional review criteria (as applicable for the project proposed). Reviewers will consider each of the review criteria below in the determination of scientific merit, and give a separate score for each. An application does not need to be strong in all categories to be judged likely to have major scientific impact. For example, a project that by its nature is not innovative may be essential to advance a field.

Specific to this FOA:

• How well has the applicant addressed potential hurdles in the development of technology for single vesicle isolation and characterization?
• Does the UG3 phase focus on technology development and feasibility using cell culture, animal models, or human biofluids?
• Does the UH3 phase include multiple human biofluids? Are the methods scalable?
• If the proposed aims of the project are fully successful, will the strategy or technology dramatically change the ability of the scientific community sort and track EV subpopulations and their exRNA cargo?

Specific to this FOA:

• Has the PD/PI or investigative team previously shown the ability to participate in, or lead, large complex projects?
• Does the PD/PI have experience in working collaboratively within a consortium or comparable groups?
• Does the PD/PI have a proven record of technology innovation??
• Does the PD/PI or investigative team have expertise in exRNA isolation in physiologically relevant human 3D cell models, tissue types derived from primary biopsies, animal models, or complex human biofluids?

Specific to this FOA?:

• Will the proposed isolation and tracking strategy/strategies improve the state of the art either by enabling isolation of EV subpopulations and cargo in a way that has previously not been possible or by dramatically improving (by at least 5-fold) our ability to purify EV subpopulations and cargo with respect to increased purity, increased speed, or decreased cost?

Specific to this FOA:

• Are the technologies or experimental approaches state of the art? Will the expected results lead to advances in technologies for single vesicle isolation and characterization?
• Are the proposed approaches, tools and technologies scientifically justified in relation to cell/biofluid source? Are the methods and procedures for characterizing and validating vesicular cargo scientifically sound and clearly explained?
• Does the application identify major technical risks, and are the proposed efforts to mitigate or address the risks clearly defined and feasible?
• Are appropriate, quantitative milestones provided for the UG3 and UH3 stages clearly defined? Are the UG3 and UH3 milestones feasible, well developed, and quantifiable with regard to the specific aims of each stage? Is the timeline feasible for the UG3 and UH3 stages? Are the critical decision points (i.e. go/no go decision points) and timelines appropriate for the UG3 and UH3 stages? Are adequate criteria provided in the UG3 stage to assess milestone completion in order to make a decision to advance studies to the UH3 stage?
• Is the proposed transition plan to the UH3 validation phase complete and in a logical sequence to the elements of the phased UG3/UH3? Do the evaluation plans, milestones and timelines
• proposed clearly identify successful completion of the UG3 and the appropriateness of advancement to the UH3 phase? Is at least one human biofluid used in the UH3 Phase?
• Are data submission, management, and support procedures described sufficiently to allow efficient and timely upload of data to the DMRR?

As applicable for the project proposed, reviewers will evaluate the following additional items while determining scientific and technical merit, and in providing an overall impact score, but will not give separate scores for these items. As applicable for the project proposed, reviewers will consider each of the following items, but will not give scores for these items, and should not consider them in providing an overall impact score.

The following special terms of award are in addition to, and not in lieu of, otherwise applicable U.S. Office of Management and Budget (OMB) administrative guidelines, U.S. Department of Health and Human Services (DHHS) grant administration regulations at 45 CFR Part 75, and other HHS, PHS, and NIH grant administration policies.

The administrative and funding instrument used for this program will be the cooperative agreement, an "assistance" mechanism (rather than an "acquisition" mechanism), in which substantial NIH programmatic involvement with the awardees is anticipated during the performance of the activities. Under the cooperative agreement, the NIH purpose is to support and stimulate the recipients' activities by involvement in and otherwise working jointly with the award recipients in a partnership role; it is not to assume direction, prime responsibility, or a dominant role in the activities. Consistent with this concept, the dominant role and prime responsibility resides with the awardees for the project as a whole, although specific tasks and activities may be shared among the awardees and the NIH as defined below.

Definitions

NIH exRNA Communication Working Group (WG): Consists of NIH programmatic staff from multiple Institutes and Centers of the NIH. This group will be primarily responsible for the stewardship of the exRNA Communication Program.

Steering Committee (SC): The SC will provide coordination activities for the exRNA Communication Program. The SC will include PDs/PIs of each of the awards and NIH exRNA Communication WG members. The SC will be chaired by two PD/PIs that are nominated by the SC and approved by the NIH. The SC will establish subcommittees to oversee the development and implementation of consortium policies including data release. The number of NIH votes may not exceed one third of the total number of votes on the SC. The number of votes from a multiple PI award will be one. Votes will inform recommendations to the NIH.

External Scientific Consultants (ESC): The NIH exRNA Communication WG will recruit outside experts (4-6 senior scientists; non-awardees) of relevance to the exRNA Communication Program. The ESC will be responsible for reviewing and evaluating the progress of the exRNA Communications Consortium toward meeting their individual and collective goals. The ESC will be appointed by and provide recommendations to the NIH about continued support of the components of the Consortium. The membership of the ESC may be enlarged permanently, or on an ad hoc basis, as needed. The ESC will meet at least once a year. During part of this meeting, there will be a joint meeting with the Steering Committee to allow the ESC to interact directly with the awardees. Annually, the ESC will provide comments regarding progress of the Consortium and on any changes that may be necessary to the NIH.

ExRNA Communication Consortium (ERCC): The ERCC will be made up of exRNA Communication awardees, the NIH exRNA Communication WG and other scientists and groups the SC agrees to include within the Consortium. The Consortium structure is meant to enable the overall goals of the exRNA Communication Program.

The PD(s)/PI(s) will have the primary responsibility for:

• Determining experimental approaches, designing protocols, setting project milestones and conducting experiments;
• Adhere to the NIH policies regarding intellectual property, data release and other policies that might be established during the course of this activity;
• Submit quarterly progress reports during the two year UG3, in a format as agreed upon by the exRNA Communication SC. Projects that are selected for continued support through the UH3 mechanism will submit progress reports on a regular basis, at least biannually;
• Accept and implement any other common guidelines and procedures developed for the NIH exRNA Communication WG and approved by the exRNA Communication SC;
• Accept and participate in the cooperative nature of the ERCC;
• Attend bi-annual workshops organized by the NIH and the Data Management and Resource Repository (DMRR).

Publications

The Program Director/Principal Investigator (PD/PI) will be responsible for the timely submission of all abstracts, manuscripts and reviews (co)authored by project investigators and supported in whole or in part under this Cooperative Agreement. The PD/PI and Project Leaders are requested to submit manuscripts to the NIH Project Scientist within two weeks of acceptance for publication so that an up-to-date summary of program accomplishments can be maintained. Publications and oral presentations of work conducted under this Cooperative Agreement are the responsibility of the Principal Investigator and appropriate Project Leaders and will require appropriate acknowledgement of NIH support. Timely publication of major findings is encouraged.

Intellectual Property

• The awardee is solely responsible for the timely acquisition of all appropriate proprietary rights, including intellectual property rights, and all materials needed for the applicant to perform the project.
• Before, during, and subsequent to the award, the U.S. Government is not required to obtain for the awardee any proprietary rights, including intellectual property rights, or any materials needed by the awardee to perform the project.
• The awardee is required to report to the U.S. Government all inventions made in the performance of the project, as specified by 35 U.S.C. Sect. 202 (Bayh-Dole Act).
• Awardees will retain custody of and have primary rights to the data and resources developed under these awards, subject to Government rights of access consistent with current HHS, PHS, and NIH policies.

NIH staff have substantial programmatic involvement that is above and beyond the normal stewardship role in awards.

NIH Project Scientists:

• Participate (with the other Steering Committee members) in the group process of setting research priorities, deciding optimal research approaches and protocol designs, and contributing to the adjustment of research protocols, project milestones or approaches as warranted. The Project Scientists will assist and facilitate the group process but not direct it;
• Serve as a liaison between the awardees, the Advisory Councils for those Institutes that plan to administer elements of the NIH Common Fund Extracellular RNA Communication Program and the larger scientific community;
• Coordinate the efforts of the awardee with others engaged in exRNA research, including other awardees under this FOA and those awardees involved in related NIH programs;
• Attend all Steering Committee meetings and assist in developing operating guidelines, quality control procedures, and consistent policies for dealing with recurrent situations that require coordinated action;
• Periodically report progress to the Directors, NIH Institutes involved in the NIH Common Fund Extracellular RNA Communication Program and the Directors of the Division of Program Coordination, Planning, and Strategic Initiatives and Office of Strategic Coordination;
• Lend relevant expertise and overall knowledge of NIH-sponsored research to facilitate the selection of scientists not affiliated with the awardee institutions who are to serve as Program Consultants.
• Serve as liaison between the Steering Committee and the Program Consultants;
• Serve on subcommittees/working groups of the Steering Committee as appropriate;
• Provide advice in the management and technical performance of the investigation;
• Assist in promoting the availability of data and resources developed in the course of this project to the scientific community at large;
• Assist awardees in the development, if needed, of policies for dealing with situations that require coordinated action;
• Retain the option to recommend the withholding or reduction of support from any cooperative agreement that either substantially fails to achieve its goals according to the milestones agreed to at the time of award, fails to maintain state-of-the-art capabilities, or fails to comply with the Terms and Conditions of the award.

Areas of Joint Responsibility include:

A SC will serve as the main governing board of the ERCC. The SC membership will include NIH Project Scientists and the PD/PI of each awarded cooperative agreement. The PD/PI of each award (or designee) will have one vote on the SC. The Project Scientists may vote, but the total votes will count as a maximum of one-third of the total number of votes. The SC Chairs will not be NIH staff members. Additional members may be added by action of the SC. Other government staff may attend the SC meetings, if their expertise is required for specific discussions. The SC will:

• Discuss progress in meeting the goals of various exRNA projects;
• Develop recommendations for uniform procedures and policies necessary to meet the goals of the Research Network, for example for data quality measures and assessment, conventions for data deposition, or measuring costs and throughput. Adoption of procedures and policies developed by the SC will require concurrence by ESC.
• The SC will also serve as a venue for coordination on the improvement of exRNA scientific methods, for example by disseminating best practices and collectively evaluating new procedures, resources, and technologies.
• The SC will consider collective goals for the ERCC, will determine how joint publication of results will contribute toward the goals of the exRNA Program, and will coordinate joint publication as needed to demonstrate overarching principles of exRNAs. It will schedule the time for, and prepare concise (2 to 4 pages) summaries of, the SC meetings, which will be delivered to members of the group within 10 business days after each meeting.
• Each full member will have one vote. Awardee members of the SC will be required to accept and implement policies approved by the SC.

Communication Plan

• Participating in regular (monthly) conference calls with all NIH exRNA Communication WG members.
• Coordinating efforts with other awardees, especially in circumstances where synergy of efforts and resources is beneficial to the overall goals of the exRNA Communication program.
• Participating and presenting findings at the semi-annual workshops convened by the NIH.
• Coordinating or jointly publishing findings in a timely manner, and as to have the broadest impact.
• Making new information and materials known to the research community in a timely manner through publications, web announcements, reports to the NIH exRNA Communication WG, and other mechanisms.

Performance Requirements

• Meeting yearly milestones as defined by investigators and NIH program at the time of award.
• Working with, cooperatively interacting with, and actively seeking input from NIH.
• Sharing broadly data and biological specimens within the Consortium and with the broader scientific community, using established data sharing guidelines , as appropriate and consistent with achieving the goals of the program.
• Attending one in-person "kickoff meeting" at the beginning of the award period; this meeting will serve as an organizing workshop together with NIH staff, including representatives from the institutes supporting the awards. The workshop will go over Terms and Conditions of Award for the UG3/UH3, ERCC responsibilities and duties, and provide an opportunity for awardees to describe their project to the consortium.
• Attending semi-annual consortium meetings to be held in the Washington, DC area.
• Attending one in-person closeout meeting at the end of the award period; this meeting will serve as a summary of accomplishments completed in the project period.
• The NIH will enlist additional scientific experts as necessary from within the NIH, other government agencies, such as the FDA and NIST whose function will be to assist the NIH exRNA Communication WG in carrying out the goals and aims of the approved studies.
• Consistent with recent NIH guidance on rigor and reproducibility in research (https://grants.nih.gov/reproducibility/index.htm), the most important milestone for the transition will be independent validation of any separation tools/technologies/approaches. To achieve this milestone, each applicant must set-aside approximately 10% (Direct Costs) of year 2-4 funds to establish a collaboration(s) with a group(s) who will independently validate and provide an assessment of the potential utility of the tools/technologies/approaches.

Dispute Resolution:

Any disagreements that may arise in scientific or programmatic matters (within the scope of the award) between award recipients and the NIH may be brought to Dispute Resolution. A Dispute Resolution Panel composed of three members will be convened. It will have three members: a designee of the Steering Committee chosen without NIH staff voting, one NIH designee, and a third designee with expertise in the relevant area who is chosen by the other two; in the case of individual disagreement, the first member may be chosen by the individual awardee. This special dispute resolution procedure does not alter the awardee's right to appeal an adverse action that is otherwise appealable in accordance with PHS regulation 42 CFR Part 50, Subpart D and DHHS regulation 45 CFR Part 16. eRA Service Desk (Questions regarding ASSIST, eRA Commons, application errors and warnings, documenting system problems that threaten on-time submission, and post-submission issues)

Finding Help Online: http://grants.nih.gov/support/ (preferred method of contact)
Telephone: 301-402-7469 or 866-504-9552 (Toll Free)

General Grants Information (Questions regarding application processes and NIH grant resources)
Email: [email protected] (preferred method of contact)
Telephone: 301-945-7573

Grants.gov Customer Support (Questions regarding Grants.gov registration and Workspace)
Contact Center Telephone: 800-518-4726
Email: [email protected]

Danilo A. Tagle, Ph.D.
National Center for Advancing Translational Sciences (NCATS)
Telephone: 301-594-8064
Email: [email protected]

Kevin Howcroft, Ph.D.
National Cancer Institute (NCI)
Telephone: 240-276-6229
Email: [email protected]

Maqsood Wani
Center for Scientific Review (CSR)
Telephone: 301-435-2270
?Email: [email protected] Gavin Wilkom
National Center for Advancing Translational Sciences (NCATS)
Telephone: 301-435-0964
Email: [email protected] Awards are made under the authorization of Sections 301 and 405 of the Public Health Service Act as amended (42 USC 241 and 284) and under Federal Regulations 42 CFR Part 52 and 45 CFR Part 75.